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Suresh K.,Indian National Institute of Animal Nutrition and Physiology | Chandrashekara S.,ChanRe Rheumatology and Immunology Center and Research
Journal of Human Reproductive Sciences | Year: 2012

Determining the optimal sample size for a study assures an adequate power to detect statistical significance. Hence, it is a critical step in the design of a planned research protocol. Using too many participants in a study is expensive and exposes more number of subjects to procedure. Similarly, if study is underpowered, it will be statistically inconclusive and may make the whole protocol a failure. This paper covers the essentials in calculating power and sample size for a variety of applied study designs. Sample size computation for single group mean, survey type of studies, 2 group studies based on means and proportions or rates, correlation studies and for case-control for assessing the categorical outcome are presented in detail. Source

Ambarish V.,Ms Ramaiah Medical College | Chandrashekara S.,Chanre Rheumatology and Immunology Center | Suresh K.P.,Indian National Institute of Animal Nutrition and Physiology
Indian Journal of Physiology and Pharmacology | Year: 2012

Exercises induce pro-inflammatory cytokines. We assessed the effect of different grades of exercises on inflammatory cytokine response. Twenty healthy volunteers performed a single bout of moderate exercise, a single bout of strenuous exercise and one month regular moderate exercise using standardized 10m Shuttle Walk Test. Interleukin-6 (IL-6) and Tumour Necrosis Factor Alpha (TNF-∝) were estimated by Sandwich ELISA method after each exercise regime. Statistics were run using SPSS software version 11.0, Systat software. Repeated measures ANOVA has been used for analysis of IL-6 values and Friedman test has been used for analyzing TNF-∝ and IL-6 values. Twenty healthy volunteers (18 to 30 years) were chosen for this study. The mean and SEM of plasma levels (pg/ ml) of IL-6 before exercise was 10.70±1.11 pg/ml, whereas, after acute moderate exercise and acute strenuous exercise it was 12.00±1.09 pg/ml and 13.35±0.89 pg/ml respectively. Interestingly, after one month of moderate exercise the values decreased to; 8.80±0.65 pg/ml. Mean and SEM of TNF-α before exercise was 121.78±29.06 pg/ml. With acute moderate exercise and after acute strenuous exercise the values were 132.90±35.75 pg/ml and 112.05±29.89 pg/ml respectively. After one month moderate exercise the levels decreased to 94.95±27.29 pg/ml. The observed changes in both IL-6 and TNF-α levels before and after both moderate and strenuous exercise were statistically significant. Although there was a slight decrease in the value of both the cytokines after one month of regular moderate exercise compared to baseline value, the difference in the values was not statistically significant. However, both IL-6 and TNF- levels showed overall statistically significant difference among the different grades of exercise. Plasma IL-6 and TNF- increase with acute moderate exercise and IL-6 increases further with acute strenuous exercise. Their levels tend to fall below baseline with one month of regular moderate exercise indicating that regular moderate exercise has beneficial effects. Source

Jodar M.,Wayne State University | Selvaraju S.,Wayne State University | Selvaraju S.,Indian National Institute of Animal Nutrition and Physiology | Sendler E.,Wayne State University | And 2 more authors.
Human Reproduction Update | Year: 2013

background: Spermatozoa are highly differentiated, transcriptionally inert cells characterized by a compact nucleus with minimal cytoplasm. Nevertheless they contain a suite of unique RNAs that are delivered to oocyte upon fertilization. They are likely integrated as part of many different processes including genome recognition, consolidation-confrontation, early embryonic development and epigenetic transgenerational inherence. Spermatozoal RNAs also provide a window into the developmental history of each sperm thereby providing biomarkers of fertility and pregnancy outcome which are being intensely studied. methods: Literature searches were performed to review the majority of spermatozoal RNA studies that described potential functions and clinical applications with emphasis on Next-Generation Sequencing. Human, mouse, bovine and stallion were compared as their distribution and composition of spermatozoal RNAs, using these techniques, have been described. results: Comparisons highlighted the complexity of the population of spermatozoal RNAs that comprises rRNA,mRNAand both large and small non-coding RNAs. RNA-seq analysis has revealed that only a fraction of the larger RNAs retain their structure. While rRNAs are the most abundant and are highly fragmented, ensuring a translationally quiescent state, other RNAs including some mRNAs retain their functional potential, thereby increasing the opportunity for regulatory interactions. Abundant small non-coding RNAs retained in spermatozoa include miRNAs and piRNAs. Some, like miR-34c are essential to the early embryo development required for the first cellular division. Others like the piRNAs are likely part of the genomic dance of confrontation and consolidation. Other non-coding spermatozoal RNAs include transposable elements, annotated lnc-RNAs, intronic retained elements, exonic elements, chromatin-associated RNAs, small-nuclear ILF3/NF30 associated RNAs, quiescent RNAs, mse-tRNAs and YRNAs. Some non-coding RNAs are known to act as epigenetic modifiers, inducing histone modifications and DNA methylation, perhaps playing a role in transgenerational epigenetic inherence. Transcript profiling holds considerable potential for the discovery of fertility biomarkers for both agriculture and human medicine. Comparing the differential RNA profiles of infertile and fertile individuals as well as assessing species similarities, should resolve the regulatory pathways contributing to male factor infertility. conclusions: Dad delivers a complex population of RNAs to the oocyte at fertilization that likely influences fertilization, embryo development, the phenotype of the offspring and possibly future generations. Development is continuing on the use of spermatozoal RNA profiles as phenotypic markers of male factor status for use as clinical diagnostics of the father's contribution to the birth of a healthy child. © The Author 2013. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. Source

Suresh K.,Indian National Institute of Animal Nutrition and Physiology
Journal of Human Reproductive Sciences | Year: 2011

Randomization as a method of experimental control has been extensively used in human clinical trials and other biological experiments. It prevents the selection bias and insures against the accidental bias. It produces the comparable groups and eliminates the source of bias in treatment assignments. Finally, it permits the use of probability theory to express the likelihood of chance as a source for the difference of end outcome. This paper discusses the different methods of randomization and use of online statistical computing web programming (www.graphpad.com/quickcalcs or www.randomization.com) to generate the randomization schedule. Issues related to randomization are also discussed in this paper. Source

Roy S.C.,Indian National Institute of Animal Nutrition and Physiology | Ghosh J.,Indian National Institute of Animal Nutrition and Physiology
Molecular Reproduction and Development | Year: 2010

In ruminants, the phenomenon of endometrial tissue remodeling during the estrous cycle and early pregnancy is not fully understood. In this report, the occurrence of tissue remodeling, if any, in buffalo endometrium was studied by detecting gelatinases, matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs); the key regulators of tissue remodeling, in uterine luminal fluids (ULF) of cycling and early pregnant (approx. 43-65 days) buffaloes. Each stage of the estrous cycle and pregnant ULF demonstrated a unique profile of gelatinase activities compared to serum/follicular fluid, with a major gelatinase band of 60 kDa with highest activity in early-luteal stage. In addition to a 32 kDa uterus-specific gelatinase band detected in both non-pregnant and pregnant ULFs, the pregnant ULF displayed three new gelatinase bands of 86, 78, and 57 kDa. Western blot technique confirmed the presence of MMP-2 (54 kDa), MMP-9 (76/73 kDa), TIMP-1 (32 kDa), TIMP-2(20 kDa), and two molecular weight forms (31 and 22 kDa) of TIMP-3 in buffalo ULF with varying band intensities. Highest MMP-2 and MMP-9 activities were observed in follicular and early-luteal stage ULFs, respectively. Highest TIMP-1 activity was observed in early-luteal ULF. Interestingly, TIMP-2 activity was only detected in mid-luteal, late-luteal, and follicular stage ULFs with significantly increasing intensities. Highest activities of 31 and 22 kDa TIMP-3 were associated with late-luteal and early-luteal stage ULFs, respectively. The varied activities of MMPs and TIMPs in buffalo ULF during the estrous cycle and early pregnancy might be a reflection of dynamic structural remodeling of the endometrium and/or developing conceptus. © 2010 Wiley-Liss, Inc. Source

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